Metatranscriptome of Feces
Bacterial and human ribosomal RNAs (rRNA) were removed from the samples using a combination of the reagents from RiboMinus Bacteria 2.0 Transcriptome Isolation and RiboMinus Eukaryotic Kits v2. Target probes were mixed in a 4:1 ratio of bacteria to human and used following the manufacturers protocol for the Bacteria 2.0 kit. Libraries from the purified RNAs were prepared with a NEBNext Ultra II RNA Library kit. Libraries were quantified, pooled and sequenced with the paired-end 75 base protocol on a NextSeq 550 using the High-Output flow cell.